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Příprava mutantního serpinu z klíštěte \kur{Ixodes ricinus}
EDEROVÁ, Monika
Point mutation altering arginin for tryptophan amino acid residue in P1 site of tick salivary serpin Iripin-1 was created using specific primers. Recombinant protein with this mutation in nucleotide sequence was then expressed in chemically competent Escherichia coli cells, extracted from them and purified by affinity and size-exclusion chromatography. To see the impact of the mutation on inhibitory function of Iripin-1, its ability to bind trypsin and form covalent complexes was evaluated.
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Alpha-gal - dvě strany jedné mince
ŠABATKOVÁ, Klára
Ticks are interesting ectoparasites for their ability to modulate the host's defense response. This ability of tick saliva is often exploited by pathogens to increase their transmission. Therefore many researchers focused on identification of molecules in tick saliva and their possible medical use. This thesis focuses on the high natural presence of anti-Gal antibodies in human serum and it's interaction with alpha-gal epitope that is expressed on the surface of some pathogens and mammalian cells. This epitope has been identified in tick saliva and some drugs as well.
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Anti-inflammatory and immunomodulatory activities of Ixodes ricinus salivary proteins
CHLASTÁKOVÁ, Adéla
Ticks are highly specialized obligate ectoparasites whose life cycle is fully dependent on obtaining a blood meal from a host. To prevent blood clotting and counteract the host's immune and wound healing responses, ticks secrete saliva, containing hundreds to thousands of proteins and non-proteins, into the feeding cavity. This thesis focuses on deciphering the function of three serpins (inhibitors of serine proteases) expressed in the salivary glands of the European tick Ixodes ricinus. Iripin-1, Iripin-3, and Iripin-8 were produced in the Escherichia coli expression system, and their effects on the immune response were evaluated using various in vitro assays and a mouse model of acute inflammation. Furthermore, the thesis involves one review article that summarizes the role of serpins in tick physiology and tick-host interaction.
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Vliv klíštěcích slin na myší neutrofilní granulocyty aktivované virem klíšťové encefalitidy
VESELÁ, Dominika
Tick saliva contain molecules that suppress the immune response of host organism thereby faciliting the transmission of pathogens by tick vectors. Neutrophils are the most common type of white blood cells. They participate in the acute phase of inflammation and via excretion of cytokines they mobilize other components of the immune system. This bachelor´s thesis focused on demonstrating of tick-borne encephalitis virus replication in neutrophils and on detection of cytokine production by neutrophilic granulocytes upon activation by the tick-borne encephalitis virus and the possible impact of tick saliva of tick Ixodes ricinus on the parameters mentioned above.
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Measurement of signalling pathway activation in mouse macrophage line IC-21 and primery dendritic cells after infection with tick-borne encephalitis virus.
Kožantová, Jana ; Lieskovská, Jaroslava (advisor) ; Drbal, Karel (referee)
Tick-borne encephalitis is a serious disease of the central nervous system. It is caused by tick-borne encephalitis virus, which is transmitted by ticks. The Czech Republic is one of the countries with the highest prevalence of this disease. Tick-borne encephalitis virus is able to replicate in several cell types. In this work we focused on macrophage line IC-21 and dendritic cells, because these cells are the first, which encounter the virus and support its spreading in the host at early stage of infection. So far there is not known any specific receptor for virus entry into cells or which signaling pathways activates. Therefore, we decided to investigate the activation of selected signaling pathways after infection with tick-borne encephalitis virus and influence of tick saliva on this activation. We employed methods of dual luciferase reporter assay, immunosandwich assay and western blot. The obtained results showed that in virus infected IC-21 cells are activated phosphatidyl-inositol pathway, NF-κB pathway, signaling molecule Erk1/2 and others. Testing of tick saliva effect revealed significantly decreased activity of NF-κB, AP-1 and CREB.
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Vliv klíštěcích slin na žírné buňky na úrovni signálních drah
HEJDOVÁ, Barbora
Intracellular signalling molecules create the signalling cascades which enable the transfer of the signal to the cell. In this work we have studied the influence of tick saliva on the cytokine production and the activation of signalling pathways in ionomycin stimulated murine mast cells. We found out that tick saliva inhibits production of several cytokines and affects two important signalling pathways in mast cells possibly involved in the regulation of cytokine induction.
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